Use of an Oxygen-Insensitive Microscale Biosensor for Methane To Measure Methane Concentration Profiles in a Rice Paddy

ABSTRACT An oxygen-insensitive microscale biosensor for methane was constructed by furnishing a previously described biosensor with an oxygen guard. The guard consisted of a glass capillary containing heterotrophic bacteria, which consumed oxygen diffusing through the tip membrane, thus preventing it from diffusing into the methane-sensing unit. Oxygen microprofiles were measured through the oxygen guard capillary, demonstrating the principle and limitations of the method. When the tip of the guard capillary was exposed to 100% oxygen at 21°C, heterotrophic oxygen consumption prevented oxygen from diffusing further than 170 μm into the capillary, whereas atmospheric levels of oxygen were consumed within 50 μm. The capacity of the oxygen guard for scavenging oxygen decreased with decreasing temperature, and atmospheric levels of oxygen caused oxygen penetration to 200 μm at 5°C. The sensors could be manufactured with tip diameters as small as 25 μm, and response times were about 1 min at room temperature. Pore water profiles of methane concentrations in a rice paddy soil were measured, and a strong correlation between the depths of oxygen penetration and methane appearance was observed as a function of the light regimen; this finding confirmed the role of microbenthic photosynthesis in limiting methane emissions from surfaces of waterlogged sediments and soils.